OBJECTIVE: To purify Rauscher Leukemia Virus to homogeneity and to fully characterize the enzyme's properties regarding optimal conditions for DNA synthesis and the biochemical parameters affecting enzyme-template-primer binding. APPROACH: The enzyme will be purified to near-homogeneity using affinity chromatography on polycytidylate-agarose, a procedure which we have developed. Purification to complete homogeneity will be accomplished through the use of additional techniques such as ion-exchange chromatography or velocity gradient centrifugation. Associated nuclease activities will be examined, and polymerase binding to template-primer in the presence and absence of catalysis to determine actual binding constants and those factors affecting such binding will be examined using insolubilized template-primers. If sufficient enzyme quantities are available, monospecific antisera against the RLV polymerase will be prepared and used for the development of a sensitive radioimmune assay for the enzyme.